Intracranial Orthotopic Model: A Procedure to Implant Cancer Cells in Mouse Brain

For cancer cell injection, attach the automatic injector unit to the stereotactic apparatus and load 6 to 8 microliters of cells, thoroughly resuspended in DPBS, into a sterile Hamilton syringe.

Load the syringe onto the injector and dispense a small volume of solution onto a disposable sterile drape to prime the needle for injection.

Use a cotton tip applicator to wipe the syringe with 70% ethanol and align the needle tip to the center of the calvarial window until it nearly touches the exposed cerebrum.

Reset the digital Vernier scale to zero and slowly insert the needle into a 3-millimeter depth into the brain. Leave the needle in the brain for at least 60 seconds before selecting RUN on the injector screen to begin the cell delivery to the injection site.

When all of the cells have been delivered, allow the needle to rest in the brain for at least 3 minutes to allow the brain parenchyma to acclimate to the injection, before retracting the needle from the brain at a rate of 0.75% millimeters per minute.